The UL2 open reading frame of bovine herpesvirus 1 encodes a uracil-DNA glycosylase.

Abstract

Sequence analysis within the unique long segment of the bovine herpesvirus 1 (BHV-1) genome previously identified an open reading frame (ORF), designated UL2, whose deduced polypeptide of 204 amino acids contained a consensus uracil-DNA glycosylase (UDGase) signature sequence. To determine whether the BHV-1 UL2 ORF product has UDGase activity, we positioned the UL2 sequence down-stream of the T7 promoter on the vector pET-28b(+) and expressed it in Escherichia coli. Upon induction with isopropyl beta-D-thiogalactopyranoside these cells produced a 23-kDa protein, the molecular mass of which was in accordance with the prediction from the nucleotide sequence. A one-step purification procedure using nickel-chelating affinity chromatography resulted in a homogeneous preparation of this protein, which displayed specific UDGase activity in an in vitro enzyme assay. These results provide evidence that the BHV-1 UL2 gene does encode a UDGase.

• Authors: Y T Chung, W Hsu
• Journal: Microbiology and immunology (Microbiol Immunol) Vol. 40 Issue 12 Pg. 949-53 ( 1996) ISSN: 0385-5600 [Print] Australia
• PMID: 9013493 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Deoxyuracil Nucleotides
• Recombinant Proteins
• deoxyuridine triphosphate
• DNA Glycosylases
• N-Glycosyl Hydrolases
• Uracil-DNA Glycosidase

Topics

• Amino Acid Sequence
• Cloning, Molecular
• Consensus Sequence
• DNA Glycosylases
• Deoxyuracil Nucleotides (metabolism)
• Escherichia coli (genetics)
• Genes, Viral
• Herpesvirus 1, Bovine (enzymology, genetics)
• Molecular Weight
• N-Glycosyl Hydrolases (chemistry, genetics, isolation & purification, metabolism)
• Open Reading Frames (genetics)
• Plasmids
• Polymerase Chain Reaction
• Recombinant Proteins (chemistry, isolation & purification, metabolism)
• Uracil-DNA Glycosidase